Thanks Artem and Frank (Was: Re: Ammonium Sulfate)
klenchin at REMOVE_TO_REPLY.facstaff.wisc.edu
Wed Aug 8 18:05:47 EST 2001
"Dr. Artem Evdokimov" <eudokima at mail.ncifcrf.gov> wrote:
>1) Size exclusion
>3) Native gels at pH 6.5 and 8.5
These are the two I was thinking of but wasn't sure if they are
sufficiently meaningful. Light-scattering - what exactly does it
measure - size? or is it sensitive enough for conformation?
>If you are masochistic, you can try NMR on your sample,
No, thanks :-)
>Can you try refolding your protein around a substrate-mimic or inhibitor
Although this is third time I come accross such proteins, this time it
is actually plain ole actin. That it is soluble after denaturation/dialysis is
well-known; that it does not polymerize after this is also well-known.
I played with salts, Ca, Mg, ATP and the result is ~ same everywhere.
I was just thinking maybe it gets trapped into some conformation that
is "better" G-actin (in a sense that it is always soluble no matter what)
than a real G-actin.
Kinda fascinating problem since this is the most abundant cellular
protein in many species and the one for which, apparently, there are
special "custom-made" chaperons. The question is - why such a
complexity for a simple idea of a reversible polymerizing agent?
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